Substituted triazolobenzodiazepines

ABSTRACT

This invention relates to novel substituted triazolobenzodiazepines of the Formula I: 
                         
wherein each of the variables are defined herein and pharmaceutically acceptable salts thereof. This invention also provides compositions comprising a compound of this invention and the use of such compositions in methods of treating diseases and conditions that are beneficially treated by administering I-BET762.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 15/610,153, filed May 31, 2017 know U.S. Pat. No. 10,039,769), whichis a continuation of U.S. application Ser. No. 15/117,544, filed Aug. 9,2016 (now U.S. Pat. No. 9,694,017), which is a National Stageapplication under 35 U.S.C. § 371 of International Application No.PCT/US2015/015034, filed Feb. 9, 2015, which claims the benefit of U.S.Provisional Application Ser. No. 61/937,701, filed Feb. 10, 2014. Thedisclosures of the prior applications are considered part of and areincorporated by reference in the disclosure of this application.

BACKGROUND OF THE INVENTION

Many current medicines suffer from poor absorption, distribution,metabolism and/or excretion (ADME) properties that prevent their wideruse or limit their use in certain indications. Poor ADME properties arealso a major reason for the failure of drug candidates in clinicaltrials. While formulation technologies and prodrug strategies can beemployed in some cases to improve certain ADME properties, theseapproaches often fail to address the underlying ADME problems that existfor many drugs and drug candidates. One such problem is rapid metabolismthat causes a number of drugs, which otherwise would be highly effectivein treating a disease, to be cleared too rapidly from the body. Apossible solution to rapid drug clearance is frequent or high dosing toattain a sufficiently high plasma level of drug. This, however,introduces a number of potential treatment problems such as poor patientcompliance with the dosing regimen, side effects that become more acutewith higher doses, and increased cost of treatment. A rapidlymetabolized drug may also expose patients to undesirable toxic orreactive metabolites.

Another ADME limitation that affects many medicines is the formation oftoxic or biologically reactive metabolites. As a result, some patientsreceiving the drug may experience toxicities, or the safe dosing of suchdrugs may be limited such that patients receive a suboptimal amount ofthe active agent. In certain cases, modifying dosing intervals orformulation approaches can help to reduce clinical adverse effects, butoften the formation of such undesirable metabolites is intrinsic to themetabolism of the compound.

In some select cases, a metabolic inhibitor will be co-administered witha drug that is cleared too rapidly. Such is the case with the proteaseinhibitor class of drugs that are used to treat HIV infection. The FDArecommends that these drugs be co-dosed with ritonavir, an inhibitor ofcytochrome P450 enzyme 3A4 (CYP3A4), the enzyme typically responsiblefor their metabolism (see Kempf, D. J. et al., Antimicrobial agents andchemotherapy, 1997, 41(3): 654-60). Ritonavir, however, causes adverseeffects and adds to the pill burden for HIV patients who must alreadytake a combination of different drugs. Similarly, the CYP2D6 inhibitorquinidine has been added to dextromethorphan for the purpose of reducingrapid CYP2D6 metabolism of dextromethorphan in a treatment ofpseudobulbar affect. Quinidine, however, has unwanted side effects thatgreatly limit its use in potential combination therapy (see Wang, L etal., Clinical Pharmacology and Therapeutics, 1994, 56(6 Pt 1): 659-67;and FDA label for quinidine at www.accessdata.fda.gov).

In general, combining drugs with cytochrome P450 inhibitors is not asatisfactory strategy for decreasing drug clearance. The inhibition of aCYP enzyme's activity can affect the metabolism and clearance of otherdrugs metabolized by that same enzyme. CYP inhibition can cause otherdrugs to accumulate in the body to toxic levels.

A potentially attractive strategy for improving a drug's metabolicproperties is deuterium modification. In this approach, one attempts toslow the CYP-mediated metabolism of a drug or to reduce the formation ofundesirable metabolites by replacing one or more hydrogen atoms withdeuterium atoms. Deuterium is a safe, stable, non-radioactive isotope ofhydrogen. Compared to hydrogen, deuterium forms stronger bonds withcarbon. In select cases, the increased bond strength imparted bydeuterium can positively impact the ADME properties of a drug, creatingthe potential for improved drug efficacy, safety, and/or tolerability.At the same time, because the size and shape of deuterium areessentially identical to those of hydrogen, replacement of hydrogen bydeuterium would not be expected to affect the biochemical potency andselectivity of the drug as compared to the original chemical entity thatcontains only hydrogen.

The effects of deuterium substitution on drug metabolism are variableand unpredictable. See, for example, Fisher, M B et al, Curr Opin DrugDiscov Devel, 2006, 9:101-09 (“Fisher”). For some compounds deuterationcauses decreased metabolic clearance in vivo. For others, there is nochange in metabolism. Still others demonstrate increased metabolicclearance. The variability in deuterium effects has led experts toquestion or dismiss deuterium modification as a viable drug designstrategy for inhibiting adverse metabolism (see Foster at p. 35 andFisher at p. 101).

The effects of deuterium modification on a drug's metabolic propertiesare not predictable even when deuterium atoms are incorporated at knownsites of metabolism. Only by actually preparing and testing a deuterateddrug can one determine if and how the rate of metabolism will differfrom that of its non-deuterated counterpart. See, for example, Fukuto etal. (J. Med. Chem. 1991, 34, 2871-76). Many drugs have multiple siteswhere metabolism is possible. The site(s) where deuterium substitutionis required and the extent of deuteration necessary to see an effect onmetabolism, if any, will be different for each drug.

I-BET762, also known as2-[6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepin-4(S)-yl]-N-ethylacetamide,is known to inhibit bromodomain-containing proteins 2 (Brd2), 3 (Brd3)and 4 (Brd4) that induce the expression and production of apolipoproteinA-1 (ApoA-1) in human hepatocyte Hep G2 cells. Bromodomains are proteindomains that bind and recognize histone acetylation. Six families ofbromodomain containing proteins are known. These bromodomain-containingproteins monitor histone acetylation and regulate epigenicallycontrolled processes such as chromatin remodeling and genetranscription. The BET family includes the proteins—Brd2, Brd3 and Brd4which are potential cancer targets. Compounds such as I-BET762 can beused for the treatment of neoplasia, acute and chronic inflammatorydisease, autoimmune disorders, obesity, fatty liver, diabetes,atherosclerosis, arterial stent occlusion, heart failure, cachexia,graft versus host disease, infectious diseases associated withbromodomains, treatment of parasites, malaria, trypanosomes and forreducing male fertility. I-BET762 is currently undergoing clinicalevaluation for NUT midline carcinoma or NMC (NUT refers to the nuclearprotein in testis) and for hematologic cancer.

Despite the beneficial activities of I-BET762, there is a continuingneed for new compounds to treat the aforementioned diseases andconditions.

SUMMARY OF THE INVENTION

This invention relates to novel substituted triazolobenzodiazepines, andpharmaceutically acceptable salts thereof. This invention also providescompositions comprising a compound of this invention and the use of suchcompositions in methods of treating diseases and conditions that arebeneficially treated by administering inhibitors of bromodomains,particularly inhibitors of BET bromodomains.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The term “treat” means decrease, suppress, attenuate, diminish, arrest,or stabilize the development or progression of a disease (e.g., adisease or disorder delineated herein), lessen the severity of thedisease or improve the symptoms associated with the disease.

“Disease” means any condition or disorder that damages or interfereswith the normal function of a cell, tissue, or organ.

It will be recognized that in a synthesized compound, the small amountsof deuterium that will vary depending upon the origin of chemicalmaterials used in the synthesis. Thus, preparations of I-BET762 willinherently contain deuterated isotopologues in various small amounts.Notwithstanding this variation, the abundance of deuterium in I-BET762is small and immaterial as compared to the degree of deuteriumsubstitution in compounds of this invention. See, for instance, Wada, Eet al., Seikagaku, 1994, 66:15; Gannes, L Z et al., Comp Biochem PhysiolMol Integr Physiol, 1998, 119:725.

In the compounds of this invention any atom not specifically designatedas a particular isotope is meant to represent any stable isotope of thatatom. Unless otherwise stated, when a position is designatedspecifically as “H” or “hydrogen”, the position is understood to havehydrogen at its natural abundance isotopic composition. Also unlessotherwise stated, when a position is designated specifically as “D” or“deuterium”, the position is understood to have deuterium at anabundance that is at least 3340 times greater than the natural abundanceof deuterium, which is 0.015% (i.e., at least 50.1% incorporation ofdeuterium).

The term “isotopic enrichment factor” as used herein means the ratiobetween the isotopic abundance and the natural abundance of a specifiedisotope.

In other embodiments, a compound of this invention has an isotopicenrichment factor for each designated deuterium atom of at least 3500(52.5% deuterium incorporation at each designated deuterium atom), atleast 4000 (60% deuterium incorporation), at least 4500 (67.5% deuteriumincorporation), at least 5000 (75% deuterium), at least 5500 (82.5%deuterium incorporation), at least 6000 (90% deuterium incorporation),at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97%deuterium incorporation), at least 6600 (99% deuterium incorporation),or at least 6633.3 (99.5% deuterium incorporation).

The term “isotopologue” refers to a species in which the chemicalstructure differs from a specific compound of this invention only in theisotopic composition thereof.

The term “compound,” when referring to a compound of this invention,refers to a collection of molecules having an identical chemicalstructure, except that there may be isotopic variation among theconstituent atoms of the molecules. Thus, it will be clear to those ofskill in the art that a compound represented by a particular chemicalstructure containing indicated deuterium atoms, will also contain lesseramounts of isotopologues having hydrogen atoms at one or more of thedesignated deuterium positions in that structure. The relative amount ofsuch isotopologues in a compound of this invention will depend upon anumber of factors including the isotopic purity of deuterated reagentsused to make the compound and the efficiency of incorporation ofdeuterium in the various synthesis steps used to prepare the compound.However, as set forth above the relative amount of such isotopologues intoto will be less than 49.9% of the compound. In other embodiments, therelative amount of such isotopologues in toto will be less than 47.5%,less than 40%, less than 32.5%, less than 25%, less than 17.5%, lessthan 10%, less than 5%, less than 3%, less than 1%, or less than 0.5% ofthe compound.

The invention also provides salts of the compounds of the invention.

A salt of a compound of this invention is formed between an acid and abasic group of the compound, such as an amino functional group, or abase and an acidic group of the compound, such as a carboxyl functionalgroup. According to another embodiment, the compound is apharmaceutically acceptable acid addition salt.

The term “pharmaceutically acceptable,” as used herein, refers to acomponent that is, within the scope of sound medical judgment, suitablefor use in contact with the tissues of humans and other mammals withoutundue toxicity, irritation, allergic response and the like, and arecommensurate with a reasonable benefit/risk ratio. A “pharmaceuticallyacceptable salt” means any non-toxic salt that, upon administration to arecipient, is capable of providing, either directly or indirectly, acompound of this invention. A “pharmaceutically acceptable counterion”is an ionic portion of a salt that is not toxic when released from thesalt upon administration to a recipient.

Acids commonly employed to form pharmaceutically acceptable saltsinclude inorganic acids such as hydrogen bisulfide, hydrochloric acid,hydrobromic acid, hydroiodic acid, sulfuric acid and phosphoric acid, aswell as organic acids such as para-toluenesulfonic acid, salicylic acid,tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylicacid, fumaric acid, gluconic acid, glucuronic acid, formic acid,glutamic acid, methanesulfonic acid, ethanesulfonic acid,benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonicacid, carbonic acid, succinic acid, citric acid, benzoic acid and aceticacid, as well as related inorganic and organic acids. Suchpharmaceutically acceptable salts thus include sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate,dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide,iodide, acetate, propionate, decanoate, caprylate, acrylate, formate,isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate,succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate,hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate,dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate,terephthalate, sulfonate, xylene sulfonate, phenylacetate,phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate,glycolate, maleate, tartrate, methanesulfonate, propanesulfonate,naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and othersalts. In one embodiment, pharmaceutically acceptable acid additionsalts include those formed with mineral acids such as hydrochloric acidand hydrobromic acid, and especially those formed with organic acidssuch as maleic acid.

The compounds of the present invention (e.g., compounds of Formula I),may contain an asymmetric carbon atom, for example, as the result ofdeuterium substitution or otherwise. As such, compounds of thisinvention can exist as either individual enantiomers, or mixtures of thetwo enantiomers. Accordingly, a compound of the present invention mayexist as either a racemic mixture or a scalemic mixture, or asindividual respective stereoisomers that are substantially free fromanother possible stereoisomer. The term “substantially free of otherstereoisomers” as used herein means less than 25% of otherstereoisomers, preferably less than 10% of other stereoisomers, morepreferably less than 5% of other stereoisomers and most preferably lessthan 2% of other stereoisomers are present. Methods of obtaining orsynthesizing an individual enantiomer for a given compound are known inthe art and may be applied as practicable to final compounds or tostarting material or intermediates.

Unless otherwise indicated, when a disclosed compound is named ordepicted by a structure without specifying the stereochemistry and hasone or more chiral centers, it is understood to represent all possiblestereoisomers of the compound.

The term “stable compounds,” as used herein, refers to compounds whichpossess stability sufficient to allow for their manufacture and whichmaintain the integrity of the compound for a sufficient period of timeto be useful for the purposes detailed herein (e.g., formulation intotherapeutic products, intermediates for use in production of therapeuticcompounds, isolatable or storable intermediate compounds, treating adisease or condition responsive to therapeutic agents).

“D” and “d” both refer to deuterium. “Stereoisomer” refers to bothenantiomers and diastereomers. “Tert” and “t-” each refer to tertiary.“US” refers to the United States of America.

“Substituted with deuterium” refers to the replacement of one or morehydrogen atoms with a corresponding number of deuterium atoms.

Throughout this specification, a variable may be referred to generally(e.g., “each Y¹) or may be referred to specifically (e.g., Y^(1a) orY^(1b)). Unless otherwise indicated, when a variable is referred togenerally, it is meant to include all specific embodiments of thatparticular variable.

Therapeutic Compounds

The present invention provides a compound of Formula I:

or a pharmaceutically acceptable salt,

wherein

Y^(1a), Y^(1b) and Y² are each independently selected from hydrogen anddeuterium;

R¹ and R² are each methyl and are independently substituted with 0 to 3deuterium;

R³ is ethyl and is substituted with 0 to 5 deuterium; and

if R¹ and R² are each CH₃, R³ is CH₂CH₃, and Y² is hydrogen, then atleast one of Y^(1a) and Y^(1b) is deuterium.

In one embodiment of this invention, each Y¹ is the same, such thatY^(1a) and Y^(1b) are each hydrogen or Y^(1a) and Y^(1b) are eachdeuterium. When each Y¹ is hydrogen, in one aspect Y² is deuterium andin another aspect, Y² is hydrogen. When each Y¹ is deuterium, in oneaspect Y² is deuterium and in another aspect Y² is hydrogen.

In one embodiment of this invention, R¹ and R² are each selected fromCH₃ and CD₃. In aspects of this embodiment, R¹ and R² are each CH₃, R¹and R² are each CD₃, R¹ is CD₃ and R² is CH₃, and R¹ is CH₃ and R² isCD₃. In further aspects Y^(1a) and Y^(1b) are each hydrogen and in otherfurther aspects Y^(1a) and Y^(1b) are each deuterium.

In one embodiment of this invention, R³ is selected from —CH₂CH₃,—CD₂CH₃, —CH₂CD₃ and —CD₂CD₃. In aspects of this embodiment, R³ is—CH₂CH₃, R³ is —CD₂CD₃, and R³ is —CD₂CH₃. Table 1 illustrates variousembodiments of this invention wherein each Y¹ is the same, Y² ishydrogen or deuterium, R¹ and R² are independently selected from CH₃ andCD₃ and R³ is selected from —CH₂CH₃, —CD₂CH₃, —CH₂CD₃ and —CD₂CD₃.

TABLE 1 Exemplary Embodiments of Formula I Embodiment R¹ R² R³ I-a CH₃CH₃ —CH₂CH₃ I-b CD₃ CH₃ —CH₂CH₃ I-c CH₃ CD₃ —CH₂CH₃ I-d CH₃ CH₃ —CD₂CD₃I-e CH₃ CH₃ —CD₂CH₃ I-f CH₃ CH₃ —CH₂CD₃ I-g CD₃ CD₃ —CH₂CH₃ I-h CD₃ CH₃—CD₂CD₃ I-i CH₃ CD₃ —CD₂CD₃ I-j CD₃ CH₃ —CD₂CH₃ I-k CH₃ CD₃ —CD₂CH₃ I-lCD₃ CH₃ —CH₂CD₃ I-m CH₃ CD₃ —CH₂CD₃ I-n CD₃ CD₃ —CD₂CD₃ I-o CD₃ CD₃—CH₂CD₃ I-p CD₃ CD₃ —CD₂CH₃

In one embodiment, the compound is a compound of Formula I whereinY^(1a)═Y^(1b)═H and is selected from any one of the compounds set forthin Table 2 (below):

TABLE 2 Exemplary Compounds of Formula I Compound Y² R¹ R² R³ 101 H —CH₃—CH₃ —CD₂CD₃ 102 H —CH₃ —CH₃ —CD₂CH₃ 103 H —CH₃ —CH₃ —CH₂CD₃ 104 H —CD₃—CH₃ —CH₂CH₃ 105 H —CH₃ —CD₃ —CH₂CH₃ 106 D —CH₃ —CH₃ —CH₂CH₃ 107 D —CD₃—CH₃ —CH₂CH₃ 108 D —CH₃ —CD₃ —CH₂CH₃ 109 D —CH₃ —CH₃ —CD₂CD₃ 110 D —CH₃—CH₃ —CD₂CH₃ 111 D —CH₃ —CH₃ —CH₂CD₃ 112 H —CD₃ —CD₃ —CH₂CH₃ 113 H —CD₃—CH₃ —CD₂CD₃ 114 H —CD₃ —CH₃ —CD₂CH₃ 115 H —CD₃ —CH₃ —CH₂CD₃ 116 H —CH₃—CD₃ —CD₂CD₃ 117 H —CH₃ —CD₃ —CD₂CH₃ 118 H —CH₃ —CD₃ —CH₂CD₃ 119 D —CD₃—CD₃ —CH₂CH₃ 120 D —CD₃ —CH₃ —CD₂CD₃ 121 D —CD₃ —CH₃ —CD₂CH₃ 122 D —CD₃—CH₃ —CH₂CD₃ 123 D —CH₃ —CD₃ —CD₂CD₃ 124 D —CH₃ —CD₃ —CD₂CH₃ 125 D —CH₃—CD₃ —CH₂CD₃ 126 H —CD₃ —CD₃ —CD₂CD₃ 127 H —CD₃ —CD₃ —CD₂CH₃ 128 H —CD₃—CD₃ —CH₂CD₃ 129 D —CD₃ —CD₃ —CD₂CD₃ 130 D —CD₃ —CD₃ —CD₂CH₃ 131 D —CD₃—CD₃ —CH₂CD₃or a pharmaceutically acceptable salt thereof.

In one embodiment, the compound is a compound of Formula I whereinY^(1a)═Y^(1b)=D and is selected from any one of the compounds set forthin Table 3 (below):

TABLE 3 Exemplary Compounds of Formula I Compound Y² R¹ R² R³ 200 H —CH₃—CH₃ —CH₂CH₃ 201 H —CH₃ —CH₃ —CD₂CD₃ 202 H —CH₃ —CH₃ —CD₂CH₃ 203 H —CH₃—CH₃ —CH₂CD₃ 204 H —CD₃ —CH₃ —CH₂CH₃ 205 H —CH₃ —CD₃ —CH₂CH₃ 206 D —CH₃—CH₃ —CH₂CH₃ 207 D —CD₃ —CH₃ —CH₂CH₃ 208 D —CH₃ —CD₃ —CH₂CH₃ 209 D —CH₃—CH₃ —CD₂CD₃ 210 D —CH₃ —CH₃ —CD₂CH₃ 211 D —CH₃ —CH₃ —CH₂CD₃ 212 H —CD₃—CD₃ —CH₂CH₃ 213 H —CD₃ —CH₃ —CD₂CD₃ 214 H —CD₃ —CH₃ —CD₂CH₃ 215 H —CD₃—CH₃ —CH₂CD₃ 216 H —CH₃ —CD₃ —CD₂CD₃ 217 H —CH₃ —CD₃ —CD₂CH₃ 218 H —CH₃—CD₃ —CH₂CD₃ 219 D —CD₃ —CD₃ —CH₂CH₃ 220 D —CD₃ —CH₃ —CD₂CD₃ 221 D —CD₃—CH₃ —CD₂CH₃ 222 D —CD₃ —CH₃ —CH₂CD₃ 223 D —CH₃ —CD₃ —CD₂CD₃ 224 D —CH₃—CD₃ —CD₂CH₃ 225 D —CH₃ —CD₃ —CH₂CD₃ 226 H —CD₃ —CD₃ —CD₂CD₃ 227 H —CD₃—CD₃ —CD₂CH₃ 228 H —CD₃ —CD₃ —CH₂CD₃ 229 D —CD₃ —CD₃ —CD₂CD₃ 230 D —CD₃—CD₃ —CD₂CH₃ 231 D —CD₃ —CD₃ —CH₂CD₃or a pharmaceutically acceptable salt thereof.

In another set of embodiments, any atom not designated as deuterium inany of the embodiments set forth above is present at its naturalisotopic abundance.

The synthesis of compounds of Formula I may be readily achieved bysynthetic chemists of ordinary skill by reference to the ExemplarySynthesis and Examples disclosed herein. Relevant procedures analogousto those of use for the preparation of compounds of Formula I andintermediates thereof are disclosed, for instance in US2012220573 andUS2012252781.

Such methods can be carried out utilizing corresponding deuterated andoptionally, other isotope-containing reagents and/or intermediates tosynthesize the compounds delineated herein, or invoking standardsynthetic protocols known in the art for introducing isotopic atoms to achemical structure.

Exemplary Synthesis

A convenient method for synthesizing compounds of Formula I is depictedin Scheme 1, below.

The benzophenenone, 13, can be prepared in 2 steps from theappropriately deuterated anthranilic acid, 10, by treatment with aceticanhydride followed by treatment with the Grignard reagent, 12. Couplingof 13 with the appropriately deuterated N-protected acyl chloride, 14,to provide 15 followed by N-deprotection and subsequent ring closure,can afford intermediate 16. Intermediate 16 can be prepared withdeuterium incorporation at each of Y¹ and Y² of greater than 90% andgreater than 95% by using Compound 14 with deuterium incorporated at theappropriate positions. Such intermediates can prepared from commerciallyavailable or deuterated reagents that are readily available as describedbelow. Generation of the hydrazoneamide, 18, can be accomplished throughtreatment of 16 with phosphorous pentasulfide to yield the thioamide,17, followed by the 2-step reaction with hydrazine then theappropriately deuterated acyl chloride, 23. Ring closure to affordintermediate 19 can be effected in the presence of acid. Hydrolysis ofthe ester, 19, and subsequent treatment with appropriately deuteratedamine, 24, in the presence of a coupling reagent such as HATU, can yieldcompounds of Formula I. Using commercially available reagents anddeuterated reagents that can be readily prepared by known methods,compounds of Formula I can be prepared with greater than 90% or greaterthan 95% deuterium incorporation at each position designated as D (seebelow for details).

Starting material, 10a, may be prepared as shown in Scheme 2 usingcommercially available CD₃I, with greater than or equal to 95% orgreater than or equal to 99% deuterium incorporation at each position inR¹ designated as D. For example CD₃I, with deuterium at 99% abundance,is commercially available.

Intermediate 10b, wherein R¹═CH₃, is commercially available.

The following deuterated reagents are commercially available withdeuterium at 98-99.5% abundance:

The following reagents can be prepared from commercially availablereagents:

The preparation of intermediate 24c is shown in Scheme 3, below, and isbased upon chemistry in Raffery, M. J. et al., Aus. J. Org. Chem., 1988,41(9), pp. 1477-1489. The preparation of intermediate 14a is describedin Rose, J. E. et al., J. Chem. Soc., Perkin Trans. 1, 1995, 2, pp.157-165. Intermediates 14b and 14c may be prepared in a manner analogousto that used for the preparation of 14a, using readily availabledeuterated starting materials, such as 2,2-d2-bromoethylacetate(commercially available at 98% d), d1-methanol or d1-ethanol (bothcommercially available at 98% d).

Intermediate 24c may be prepared from 2,2-d2-ethyl iodide (commerciallyavailable at 99.5% deuterium incorporation) as referenced above.

The specific approaches and compounds shown above are not intended to belimiting. The chemical structures in the schemes herein depict variablesthat are hereby defined commensurately with chemical group definitions(moieties, atoms, etc.) of the corresponding position in the compoundformulae herein, whether identified by the same variable name (i.e., R¹,R², R³, etc.) or not. The suitability of a chemical group in a compoundstructure for use in the synthesis of another compound is within theknowledge of one of ordinary skill in the art.

Additional methods of synthesizing compounds of Formula I and theirsynthetic precursors, including those within routes not explicitly shownin schemes herein, are within the means of chemists of ordinary skill inthe art. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing theapplicable compounds are known in the art and include, for example,those described in Larock R, Comprehensive Organic Transformations, VCHPublishers (1989); Greene, T W et al., Protective Groups in OrganicSynthesis, 3^(rd) Ed., John Wiley and Sons (1999); Fieser, L et al.,Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons(1994); and Paquette, L, ed., Encyclopedia of Reagents for OrganicSynthesis, John Wiley and Sons (1995) and subsequent editions thereof.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds.

Compositions

The invention also provides pyrogen-free pharmaceutical compositionscomprising an effective amount of a compound of Formula I (e.g.,including any of the formulae herein), or a pharmaceutically acceptablesalt of said compound; and a pharmaceutically acceptable carrier. Thecarrier(s) are “acceptable” in the sense of being compatible with theother ingredients of the formulation and, in the case of apharmaceutically acceptable carrier, not deleterious to the recipientthereof in an amount used in the medicament.

Pharmaceutically acceptable carriers, adjuvants and vehicles that may beused in the pharmaceutical compositions of this invention include, butare not limited to, ion exchangers, alumina, aluminum stearate,lecithin, serum proteins, such as human serum albumin, buffer substancessuch as phosphates, glycine, sorbic acid, potassium sorbate, partialglyceride mixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

If required, the solubility and bioavailability of the compounds of thepresent invention in pharmaceutical compositions may be enhanced bymethods well-known in the art. One method includes the use of lipidexcipients in the formulation. See “Oral Lipid-Based Formulations:Enhancing the Bioavailability of Poorly Water-Soluble Drugs (Drugs andthe Pharmaceutical Sciences),” David J. Hauss, ed. Informa Healthcare,2007; and “Role of Lipid Excipients in Modifying Oral and ParenteralDrug Delivery: Basic Principles and Biological Examples,” Kishor M.Wasan, ed. Wiley-Interscience, 2006.

Another known method of enhancing bioavailability is the use of anamorphous form of a compound of this invention optionally formulatedwith a poloxamer, such as LUTROL™ and PLURONIC™ (BASF Corporation), orblock copolymers of ethylene oxide and propylene oxide. See U.S. Pat.No. 7,014,866; and United States patent publications 20060094744 and20060079502.

The pharmaceutical compositions of the invention include those suitablefor oral, rectal, nasal, topical (including buccal and sublingual),vaginal or parenteral (including subcutaneous, intramuscular,intravenous and intradermal) administration. In certain embodiments, thecompound of the formulae herein is administered transdermally (e.g.,using a transdermal patch or iontophoretic techniques). Otherformulations may conveniently be presented in unit dosage form, e.g.,tablets, sustained release capsules, and in liposomes, and may beprepared by any methods well known in the art of pharmacy. See, forexample, Remington: The Science and Practice of Pharmacy, LippincottWilliams & Wilkins, Baltimore, Md. (20th ed. 2000).

Such preparative methods include the step of bringing into associationwith the molecule to be administered ingredients such as the carrierthat constitutes one or more accessory ingredients. In general, thecompositions are prepared by uniformly and intimately bringing intoassociation the active ingredients with liquid carriers, liposomes orfinely divided solid carriers, or both, and then, if necessary, shapingthe product.

In certain embodiments, the compound is administered orally.Compositions of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, sachets, or tabletseach containing a predetermined amount of the active ingredient; apowder or granules; a solution or a suspension in an aqueous liquid or anon-aqueous liquid; an oil-in-water liquid emulsion; a water-in-oilliquid emulsion; packed in liposomes; or as a bolus, etc. Soft gelatincapsules can be useful for containing such suspensions, which maybeneficially increase the rate of compound absorption.

In the case of tablets for oral use, carriers that are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried cornstarch. When aqueoussuspensions are administered orally, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweeteningand/or flavoring and/or coloring agents may be added.

Compositions suitable for oral administration include lozengescomprising the ingredients in a flavored basis, usually sucrose andacacia or tragacanth; and pastilles comprising the active ingredient inan inert basis such as gelatin and glycerin, or sucrose and acacia.

Compositions suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents. The formulations may be presented in unit-dose or multi-dosecontainers, for example, sealed ampules and vials, and may be stored ina freeze dried (lyophilized) condition requiring only the addition ofthe sterile liquid carrier, for example water for injections,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tablets.

Such injection solutions may be in the form, for example, of a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to techniques known in the art using suitabledispersing or wetting agents (such as, for example, Tween 80) andsuspending agents. The sterile injectable preparation may also be asterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example, as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that maybe employed are mannitol, water, Ringer's solution and isotonic sodiumchloride solution. In addition, sterile, fixed oils are conventionallyemployed as a solvent or suspending medium. For this purpose, any blandfixed oil may be employed including synthetic mono- or diglycerides.Fatty acids, such as oleic acid and its glyceride derivatives are usefulin the preparation of injectables, as are naturalpharmaceutically-acceptable oils, such as olive oil or castor oil,especially in their polyoxyethylated versions. These oil solutions orsuspensions may also contain a long-chain alcohol diluent or dispersant.

In another embodiment, a composition of this invention further comprisesa second therapeutic agent. The second therapeutic agent may be selectedfrom any compound or therapeutic agent known to have or thatdemonstrates advantageous properties when administered with a compoundhaving the same mechanism of action as I-BET762. Such agents includethose indicated as being useful in combination with I-BET762, includingbut not limited to, those described in US2012220573.

Preferably, the second therapeutic agent is an agent useful in thetreatment of a disease or condition selected from neoplasia,inflammatory disease, autoimmune disorders, obesity, fatty liver,diabetes, atherosclerosis, arterial stent occlusion, heart failure,cachexia, graft versus host disease, infectious diseases associated withbromodomains, parasitic infection, malaria, trypanosomes and forreducing male fertility.

In another embodiment, the invention provides separate dosage forms of acompound of this invention and one or more of any of the above-describedsecond therapeutic agents, wherein the compound and second therapeuticagent are associated with one another. The term “associated with oneanother” as used herein means that the separate dosage forms arepackaged together or otherwise attached to one another such that it isreadily apparent that the separate dosage forms are intended to be soldand administered together (within less than 24 hours of one another,consecutively or simultaneously).

In the pharmaceutical compositions of the invention, the compound of thepresent invention is present in an effective amount. As used herein, theterm “effective amount” refers to an amount which, when administered ina proper dosing regimen, is sufficient to treat the target disorder.

The interrelationship of dosages for animals and humans (based onmilligrams per meter squared of body surface) is described in Freireichet al., Cancer Chemother. Rep, 1966, 50: 219. Body surface area may beapproximately determined from height and weight of the subject. See,e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 1970,537.

In one embodiment, an effective amount of a compound of this inventionover a course of treatment can range from 0.05 to 1000 mg and may beadministered from once to four times per day. In certain aspects of thisembodiment, an effective amount of a compound of this invention canrange from 0.1 to 1000 mg, 0.1 to 500 mg, 0.1 to 50 mg, 0.5 to 500 mg,0.5 to 100 mg, 0.5 to 50 mg, 0.5 to 5 mg, 1 to 250 mg, 1 to 100 mg, 1 to10 mg, 5 to 100 mg, 5 to 50 mg and 5 to 10 mg. In certain aspects ofthis embodiment the compound may be administered three times a day,twice a day, once a day, or once every other day, over the course oftreatment.

Effective doses will also vary, as recognized by those skilled in theart, depending on the diseases treated, the severity of the disease, theroute of administration, the sex, age and general health condition ofthe subject, excipient usage, the possibility of co-usage with othertherapeutic treatments such as use of other agents and the judgment ofthe treating physician. For example, guidance for selecting an effectivedose can be determined by reference to the prescribing information forI-BET762.

For pharmaceutical compositions that comprise a second therapeuticagent, an effective amount of the second therapeutic agent is betweenabout 20% and 100% of the dosage normally utilized in a monotherapyregime using just that agent. Preferably, an effective amount is betweenabout 70% and 100% of the normal monotherapeutic dose. The normalmonotherapeutic dosages of these second therapeutic agents are wellknown in the art. See, e.g., Wells et al., eds., PharmacotherapyHandbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDRPharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition,Tarascon Publishing, Loma Linda, Calif. (2000), each of which referencesare incorporated herein by reference in their entirety.

It is expected that some of the second therapeutic agents referencedabove will act synergistically with the compounds of this invention.When this occurs, it will allow the effective dosage of the secondtherapeutic agent and/or the compound of this invention to be reducedfrom that required in a monotherapy. This has the advantage ofminimizing toxic side effects of either the second therapeutic agent ofa compound of this invention, synergistic improvements in efficacy,improved ease of administration or use and/or reduced overall expense ofcompound preparation or formulation.

Methods of Treatment

In another embodiment, the invention provides a method of bromodomaininhibition in a cell, comprising contacting a cell with one or morecompounds of Formula I herein, or a pharmaceutically acceptable saltthereof.

According to another embodiment, the invention provides a method oftreating a disease that is beneficially treated by I-BET762 in a subjectin need thereof, comprising the step of administering to the subject aneffective amount of a compound or a composition of this invention. Inone embodiment the subject is a patient in need of such treatment. Suchdiseases are well known in the art and are disclosed in, but not limitedto, the following patents and published applications: US2012220573 andUS 2012252781. Such diseases include, but are not limited to, neoplasia,acute and chronic inflammatory disease, autoimmune disorders, obesity,fatty liver, diabetes, atherosclerosis, arterial stent occlusion, heartfailure, cachexia, graft versus host disease, infectious diseasesassociated with bromodomains, parasitic infection, malaria, trypanosomesand for reducing male fertility.

In one embodiment, chronic autoimmune and inflammatory conditionsinclude conditions such as rheumatoid arthritis, osteoarthritis, acutegout, psoriasis, systemic lupus erythematosus, multiple sclerosis,inflammatory bowel disease (Crohn's disease and Ulcerative colitis),asthma, chronic obstructive airways disease, pneumonitis, myocarditis,pericarditis, myositis, eczema, dermatitis, alopecia, vitiligo, bullousskin diseases, nephritis, vasculitis, atherosclerosis, Alzheimer'sdisease, depression, retinitis, uveitis, scleritis, hepatitis,pancreatitis, primary biliary cirrhosis, sclerosing cholangitis,Addison's disease, hypophysitis, thyroiditis, type I diabetes and acuterejection of transplanted organs.

In one embodiment, acute inflammatory conditions include conditions suchas acute gout, giant cell arteritis, nephritis including lupusnephritis, vasculitis with organ involvement such as glomerulonephritis,vasculitis including giant cell arteritis, Wegener's granulomatosis,Polyarteritis nodosa, Behcet's disease, Kawasaki disease, Takayasu'sArteritis and acute rejection of transplanted organs.

In one embodiment, the invention provides a method of treatment ofdiseases or conditions which involve inflammatory responses toinfections with bacteria, viruses, fungi, parasites or their toxins,such as sepsis, sepsis syndrome, septic shock, endotoxaemia, systemicinflammatory response syndrome (SIRS), multi-organ dysfunction syndrome,toxic shock syndrome, acute lung injury, ARDS (adult respiratorydistress syndrome), acute renal failure, fulminant hepatitis, burns,acute pancreatitis, post-surgical syndromes, sarcoidosis, Herxheimerreactions, encephalitis, myelitis, meningitis, malaria, SIRS associatedwith viral infections such as influenza, herpes zoster, herpes simplexand coronavirus.

In one embodiment, the invention provides a method of treatment ofdiseases or conditions associated with ischaemia-reperfusion injury suchas myocardial infarction, cerebrovascular ischaemia (stroke), acutecoronary syndromes, renal reperfusion injury, organ transplantation,coronary artery bypass grafting, cardio-pulmonary bypass procedures andpulmonary, renal, hepatic, gastro-intestinal or peripheral limbembolism.

In one embodiment, the invention provides a method of treatment ofdisorders of lipid metabolism via the regulation of APO-A1 such ashypercholesterolemia, atherosclerosis and Alzheimer's disease.

In one embodiment, the invention provides a method of treatment offibrotic conditions such as idiopathic pulmonary fibrosis, renalfibrosis, post-operative stricture, keloid formation, scleroderma andcardiac fibrosis.

In one embodiment, the invention provides a method of treatment of viralinfections such as herpes virus, human papilloma virus, adenovirus,poxvirus and other DNA viruses.

In one embodiment, the invention provides a method of treatment ofcancer, including hematological, epithelial including lung, breast andcolon carcinomas, midline carcinomas, mesenchymal, hepatic, renal andneurological tumors.

In one embodiment, the invention provides a method of treating a diseaseassociated with systemic inflammatory response syndrome, such as sepsis,sepsis syndrome, septic shock, endotoxaemia, burns, pancreatitis, acutepancreatitis, chronic pancreatitis, major trauma, haemorrhage andischaemia.

In one embodiment, the invention provides a method of treating a diseaseor condition selected from herpes simplex infections and reactivations,cold sores, herpes zoster infections and reactivations, chickenpox,shingles, human papilloma virus, cervical neoplasia, adenovirusinfections, including acute respiratory disease, poxvirus infectionssuch as cowpox and smallpox and African swine fever virus, and humanpapilloma virus infections of skin or cervical epithelia.

In one embodiment, the invention provides a method of treatment ofcancer, such as midline carcinoma.

In one embodiment, this invention provides a method of treatment of adisease or condition selected from carcinoma and hematologic cancer in asubject in need thereof.

In one embodiment, the method of this invention is used to treat adisease or condition selected from chronic autoimmune disease andchronic inflammatory disease in a subject in need thereof.

Identifying a subject in need of such treatment can be in the judgmentof a subject or a health care professional and can be subjective (e.g.opinion) or objective (e.g. measurable by a test or diagnostic method).

In another embodiment, any of the above methods of treatment comprisesthe further step of co-administering to the subject in need thereof oneor more second therapeutic agents. The choice of second therapeuticagent may be made from any second therapeutic agent known to be usefulfor co-administration with I-BET762. The choice of second therapeuticagent is also dependent upon the particular disease or condition to betreated. Examples of second therapeutic agents that may be employed inthe methods of this invention are those set forth above for use incombination compositions comprising a compound of this invention and asecond therapeutic agent.

The term “co-administered” as used herein means that the secondtherapeutic agent may be administered together with a compound of thisinvention as part of a single dosage form (such as a composition of thisinvention comprising a compound of the invention and an secondtherapeutic agent as described above) or as separate, multiple dosageforms. Alternatively, the additional agent may be administered prior to,consecutively with, or following the administration of a compound ofthis invention. In such combination therapy treatment, both thecompounds of this invention and the second therapeutic agent(s) areadministered by conventional methods. The administration of acomposition of this invention, comprising both a compound of theinvention and a second therapeutic agent, to a subject does not precludethe separate administration of that same therapeutic agent, any othersecond therapeutic agent or any compound of this invention to saidsubject at another time during a course of treatment.

Effective amounts of these second therapeutic agents are well known tothose skilled in the art and guidance for dosing may be found in patentsand published patent applications referenced herein, as well as in Wellset al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange,Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000),and other medical texts. However, it is well within the skilledartisan's purview to determine the second therapeutic agent's optimaleffective-amount range.

In one embodiment of the invention, where a second therapeutic agent isadministered to a subject, the effective amount of the compound of thisinvention is less than its effective amount would be where the secondtherapeutic agent is not administered. In another embodiment, theeffective amount of the second therapeutic agent is less than itseffective amount would be where the compound of this invention is notadministered. In this way, undesired side effects associated with highdoses of either agent may be minimized. Other potential advantages(including without limitation improved dosing regimens and/or reduceddrug cost) will be apparent to those of skill in the art.

In yet another aspect, the invention provides the use of a compound ofFormula I alone or together with one or more of the above-describedsecond therapeutic agents in the manufacture of a medicament, either asa single composition or as separate dosage forms, for treatment in asubject of a disease, disorder or symptom set forth above. Anotheraspect of the invention is a compound of Formula I for use in thetreatment in a subject of a disease, disorder or symptom thereofdelineated herein.

EXAMPLES Example 1.(S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d3)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 104)

Step 1. Methyl 5-hydroxy-2-nitrobenzoate (20)

Thionyl chloride (13.0 g, 109 mmol) was added slowly at room temperatureto a solution of 3-hydroxy-5-nitrobenzoic acid (20′) (20.0 g, 109 mmol)in methanol (500 mL). The reaction was heated at reflux for 3 hours,cooled to room temperature and concentrated under reduced pressure. Theresidue was dissolved in ethyl acetate (200 mL), washed with saturatedsodium bicarbonate (200 mL) and saturated sodium chloride (200 mL). Theorganic layer was dried over sodium sulfate, filtered and concentratedto afford (20) as a white solid (16 g, 74%).

Step 2. Methyl 5-(methoxy-d₃)-2-nitrobenzoate (21)

To a solution of 20 (16 g, 81.2 mmol) in anhydrous DMF (500 mL) wasadded potassium carbonate (22.4 g, 162.4 mmol), followed by methyliodide-d₃ (22a), (Cambridge Isotope, 99.5% D) (15.8 g, 106 mmol). Thereaction was stirred for 24 hours, cooled to 0° C. and water (3 L) wasadded slowly over 1 hour. The mixture was extracted with ethyl acetate(3×500 mL). The organic layer was washed with saturated sodiumbicarbonate (500 mL), water (2×500 mL), saturated sodium chloride (500mL), dried over sodium sulfate, filtered and concentrated under reducedpressure. The residue was triturated with 10% ethyl acetate in heptanesto afford (21) as an off-white solid (15 g, 86%).

Step 3. 2-Amino-5-(methoxy-d₃) benzoic acid (10a)

To a solution of 21 (15 g, 70.1 mmol) in THF (200 mL) was added 1Nsodium hydroxide (210 mL, 210 mmol). The reaction was stirred 4 hours atroom temperature, cooled to 0° C. and adjusted to pH 2 with 3Nhydrochloric acid. The resulting mixture was extracted withdichloromethane (3×200 mL) and combined organic phase was dried oversodium sulfate, filtered and concentrated to give the free acid (15 g).The acid was hydrogenated with 10% palladium on carbon (1.3 g; 50 wt %water) in methanol (300 mL) at 35 psi for 6 hours at room temperature.The mixture was filtered through a pad of celite and the filtrate wasconcentrated to afford 10a as a tan solid (10 g, 85%).

Step 4. 6-(Methoxy-d₃)-2-methyl-4H-benzo[d][1,3]oxazin-4-one (11a)

Compound (10a) (10 g, 60 mmol) was dissolved in acetic anhydride (60 mL)and heated at reflux for 6 hours and concentrated under reducedpressure. The residue was co-evaporated with toluene (3×50 mL), andsuspended in MTBE (100 mL), stirred for 15 minutes, and filtered toafford 11a as a brown solid which was taken directly to the next step(9.5 g, 79%).

Step 5. (2-Amino-5-(methoxy-d₃)phenyl)(4-chlorophenyl)methanone (13a)

To a solution of 11a (9.5 g, 49 mmol) in a mixture of toluene anddiethyl ether (3:1) (120 mL) at 0° C. was added a solution of 1.0 M4-chlorophenylmagnesium bromide (12) in methylTHF (40 mL, 40 mmol) over30 minutes. The reaction was stirred at room temperature for 1 hour,cooled to 0° C. and 1N HCl (50 mL) added. The aqueous layer wasextracted with ethyl acetate (2×100 mL), and the organic layer waswashed with saturated sodium chloride (100 mL), dried over sodiumsulfate, filtered and concentrated under reduced pressure. The residualmaterial was dissolved in ethanol (100 mL) and 6N HCl (30 mL) was added,heated at reflux for 2 hours then concentrated under reduced pressure.The residue was suspended in ethyl acetate (200 mL) and adjusted with 1NNaOH to pH 8-9. The mixture was extracted with ethyl acetate (3×100 mL)and organic layer, washed with saturated sodium chloride (2×200 mL),dried over sodium sulfate and concentrated to afford 13a as a yellowsolid (7 g, 54%).

Step 6. Methyl(S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((2-(4-chlorobenzoyl)-4-(methoxy-d₃)phenyl)amino)-4-oxobutanoate(15a)

To a solution of Fmoc-Asp (OMe)-OH (10.7 g, 29.2 mmol) indichloromethane (20 mL) was added thionyl chloride (20 mL), carefullyfollowed by dimethylformamide (0.25 mL, 3.25 mmol). The mixture wasstirred at room temperature for 3 hours, concentrated under reducedpressure and the residual material was co-evaporated with toluene (3×100mL) to give crude 14 (12.3 g). The crude solid was suspended indichloromethane (100 mL) and 13a (7 g, 26.5 mmol) was added. Thereaction was stirred at 60° C. for 2 hours and concentrated to afford15a as a white solid (14 g, 86%).

Step 7. Methyl(S)-2-(5-(4-chlorophenyl)-7-(methoxy-d₃)-2-oxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)acetate(16a)

To a solution of 15a (14 g, 22.7 mmol) in dichloromethane (50 mL) wasadded triethylamine (57 mL, 409 mmol). The reaction was heated at refluxovernight, concentrated under reduced pressure and the residual materialwas suspended in 1, 2-dichloroethane (130 mL), and acetic acid (13 mL,227 mmol) was added. The reaction was stirred at 60° C. for 2 hours,concentrated under reduced pressure and the residue was suspended indichloromethane (300 mL), washed with 1N HCl (100 mL), water (2×100 mL),saturated sodium chloride (100 mL), dried over sodium sulfate, filtered,and concentrated under reduced pressure. The residual material wassuspended in acetonitrile (15 mL), stirred for 45 minutes, and theresulting solid was filtered, dried in a vacuum oven at 40° C. overnightto afford 16a as a light brown solid (7.4 g, 87%).

Step 8. Methyl(S)-2-(5-(4-chlorophenyl)-7-(methoxy-d₃)-2-thioxo-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)acetate(17a)

Phosphorus pentasulfide (11 g, 24 mmol) was suspended in 1,2-dichloroethane (150 mL) and sodium carbonate (2.6 g, 24 mmol) wasadded. The resulting mixture was stirred at room temperature for 1 hourand 16a (5 g, 13.4 mmol) was added. The reaction was stirred at 65° C.for 4 hours, cooled to ambient temperature and filtered. The filtratewas washed with saturated sodium bicarbonate (100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The crude material was purified using anAnalogix automated chromatography system, eluting with a gradient ofethyl acetate in heptanes (0 to 30%). Product fractions were pooled andevaporated yielding 17a as a light yellow solid (3.2 g, 61.2%).

Step 9. Methyl(S,Z)-2-(2-(2-acetylhydrazono)-5-(4-chlorophenyl)-7-(methoxy-d₃)-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)acetate(18a)

To a solution of 17a (1.5 g, 3.85 mmol) in THF (60 mL), at 0° C. wasadded Hydrazine monohydrate (0.56 mL, 12.5 mmol) and the reactionstirred at 0° C. for 4 hours. Triethylamine (1.6 mL, 13.5 mmol) wasadded, followed by acetyl chloride 23, (1.64 mL, 12.5 mmol) and thereaction was stirred at room temperature for 1 hour, diluted with water(50 mL) and THF was removed under reduced pressure. The resultingresidue was extracted with dichloromethane (100 mL) and the organiclayer was washed with saturated sodium chloride (100 mL), dried oversodium sulfate, filtered, and concentrated to afford 18a as a yellowsolid (1.6 g, 96%).

Step 10. Methyl(S)-2-(6-(4-chlorophenyl)-8-(methoxy-d3)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetate(19a)

To a solution of 18a (1.6 g, 3.7 mmol) in THF (50 mL) was added aceticacid (50 mL) and the reaction was stirred at room temperature for 18hours. The mixture was concentrated under reduced pressure to afford aresidue which was dissolved in dichloromethane (100 mL), washed withsaturated sodium bicarbonate (3×50 mL), water (100 mL), saturated sodiumchloride (100 mL), and dried over sodium sulfate, filtered, andconcentrated under reduced pressure. The crude material was purifiedusing an Analogix automated chromatography system, eluting with methanolin dichloromethane (0 to 5%). Product fractions were pooled andevaporated to afford 19a as a light yellow solid (1.2 g, 82%).

Step 11.(S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d₃)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 104)

To a solution of 19a (1.2 g, 2.9 mmol) in THF (20 mL) was added 1N NaOH(8.7 mL, 8.7 mmol). The reaction was stirred for 6 hours at roomtemperature, cooled to 0° C. and adjusted with 1N HCl to pH 4. Theresulting mixture was extracted with dichloromethane (3×50 mL) and thecombined organic layer was dried over sodium sulfate, filtered, andconcentrated to give the free acid as a yellow solid (1.0 g, 86%). To asolution of the free acid (500 mg, 1.25 mmol) in THF (20 mL) was addedHATU (950 mg, 2.5 mmol), followed by diisopropylethylamine (0.42 mL, 2.5mmol) and the reaction was stirred at room temperature for 3 hours.Ethylamine hydrochloride (24) (200 mg, 2.5 mmol) was added, followed bydiisopropylethylamine (0.42 mL, 2.5 mmol) and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered and concentratedunder reduced pressure. The residual material was purified using anAnalogix automated chromatography system eluting with methanol indichloromethane (0 to 5%). Product fractions were pooled and evaporatedto give a material with enantiomeric purity of 94% ee (400 mg). Thematerial was further purified using chiral preparative HPLC (20×250 mm,10 μm column, Daicel ChiralPak AD, eluting with 80% isopropanol/20%hexane at a flow rate of 17 mL/min). Product fractions were pooled andevaporated to afford 104 as an off white solid (280 mg, 53%). ¹H-NMR(300 MHz, CDCl₃): δ 1.18 (m, 3H), 2.61 (s, 3H), 3.24-3.35 (m, 2H),3.36-3.41 (m, 1H), 3.47-3.54 (m, 1H), 4.61 (t, J=7 Hz, 1H), 6.4 (bm,1H), 6.85 (m, 1H), 7.17-7.21 (m, 1H), 7.32-7.35 (m, 2H), 7.36 (s, 1H),7.46-7.51 (m, 2H); ¹³C-NMR (75 MHz, CDCl₃): δ 12.12, 14.78, 34.49,39.47, 53.93, 115.79, 117.93, 124.81, 126.44, 128.49, 130.12, 130.73,137.94, 137.14, 150.46, 156.43, 157.98, 166.20, 170.3. MS(ESI) [(M+H)⁺]C₂₂H₁₉D₃ClN₅O₂: 427; HPLC (method: SorbTech C18AQ, 2.1×50 mm 3 μmcolumn-gradient method 5-95% ACN+0.1% formic acid in 14 min with 4 minhold at 95% ACN+0.1% formic acid; Wavelength: 254 nm): retention time:6.33 min; 99.1% purity; Chiral HPLC (method: 25 cm×4.6 mm, 10 μm columnChiralpak AD, isocratic method 40% heptane +60% EtOH for 30 minutes at1.0 mL/min, Wavelength: 210 nm): retention time: 4.654 min, purity:97.87% ee.

Example 2.S)-2-(6-(4-Chlorophenyl)-8-methoxy-1-(methyl-d3)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 105)

Step 1. Methyl(S,Z)-2-(2-(2-(acetyl-d₃)hydrazono)-5-(4-chlorophenyl)-7-methoxy-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)acetate(18b)

To a solution of 17 (1 g, 2.6 mmol) in THF (50 mL) at 0° C. was addedHydrazine monohydrate (0.38 mL, 8 mmol) and the reaction was stirred at0° C. for 4 hours. Triethylamine (1.1 mL, 7.8 mmol) was added, followedby the addition of acetyl chloride-d₃ (23a) (0.6 mL, 8 mmol, CambridgeIsotopes, 98% D) and the reaction was stirred at room temperature for 1hour, diluted with water (50 mL) and the THF was removed under reducedpressure. The residual material was extracted with dichloromethane(1×100 mL), washed with saturated sodium chloride (100 mL), dried oversodium sulfate, filtered, and concentrated to afford 18b as a brownsolid (1.1 g, 99%).

Step 2. Methyl(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)acetate(19b)

To a solution of 18b (1.1 g, 2.55 mmol) in THF (40 mL) was added aceticacid (40 mL) and the reaction was stirred at room temperature for 18hours. The resulting mixture was concentrated under reduced pressure toafford a residue which was dissolved in dichloromethane (150 mL), washedwith saturated sodium bicarbonate (3×50 mL), water (100 mL), saturatedsodium chloride (100 mL), and dried over sodium sulfate, filtered, andconcentrated under reduced pressure. The crude material was purifiedusing an Analogix automated chromatography system, eluting with methanolin dichloromethane (0 to 5%). Product fractions were pooled andevaporated to afford 19b as a light yellow solid (0.96 g, 91%).

Step 3.(S)-2-(6-(4-Chlorophenyl)-8-methoxy-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 105)

To a solution of 19b (0.96 g, 2.33 mmol) in THF (20 mL) was added 1NNaOH (7 mL, 7 mmol) and the reaction was stirred for 5 hours at 40° C.,cooled to 0° C. and adjusted with 1N HCl solution to pH 4-5. Theresulting mixture was extracted with dichloromethane (3×50 mL), and thecombined organic layer was dried over sodium sulfate, filtered, andconcentrated to give the free acid as a yellow solid (940 mg, 100%). Toa solution of the free acid (460 mg, 1.15 mmol) in THF (20 mL) was addedHATU (880 mg, 2.31 mmol), followed by diisopropylethylamine (0.4 mL,2.31 mmol) and the reaction was stirred at room temperature for 3 hours.Ethylamine hydrochloride (24) (188 mg, 2.31 mmol) was added, followed bydiisopropylethylamine (0.4 mL, 2.31 mmol) and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered and concentratedunder reduced pressure. The residual material was purified using anAnalogix automated chromatography system eluting with methanol indichloromethane (0 to 5%). Product fractions were pooled and evaporatedto give a material with enantiomeric purity of 88% ee (380 mg). Thematerial was further purified using chiral preparative HPLC (20×250 mm,10 μm column, Daicel ChiralPak AD, eluting with 80% isopropanol/20%hexane at a flow rate of 17 mL/min). Product fractions were pooled andevaporated to afford (105) as an off white solid (240 mg, 49%). ¹H-NMR(300 MHz, CDCl₃): δ 1.18 (t, J=7.3 Hz, 3H), 3.23-3.34 (m, 2H),3.35-3.3.55 (m, 2H), 3.8 (s, 3H), 4.62 (t, J=7 Hz, 1H), 6.43 (bm, 1H),6.85 (d, J=2.7 Hz, 1H), 7.18-7.22 (m, 1H), 7.27-7.34 (m, 2H), 7.35, (s,1H), 7.47-7.51 (m, 2H); ¹³C-NMR (75 MHz, CDCl₃): δ 14.78, 34.49, 39.46,53.92, 55.86, 115.81, 117.94, 124.78, 126.47, 128.49, 130.10, 130.74,136.94, 137.15, 156.43, 157.97, 166.18, 170.30. MS(ESI) [(M+H)⁺]C₂₂H₁₉D₃ClN₅O₂: 427; HPLC (method: SorbTech C18AQ, 2.1×50 mm 3 μmcolumn-gradient method 5-95% ACN+0.1% formic acid in 14 min with 4 minhold at 95% ACN+0.1% formic acid; Wavelength: 254 nm): retention time:6.353 min; 98% purity; Chiral HPLC (method: 25 cm×4.6 mm, 10 μm columnChiralpak AD, isocratic method 40% heptane +60% EtOH for 30 minutes at1.0 mL/min, Wavelength: 210 nm): retention time: 4.650 min, purity:96.91% ee.

Example 3.(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d5)acetamide(Compound 101)

(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-methyl-4H-benzo[fl][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d₅)acetamide(Compound 101)

To a solution of the free acid of 19 (400 mg, 1.01 mmol) (prepared bytreating with 1N NaOH/THF as described in step 3 of 105) was dissolvedin THF (20 mL) and HATU (768 mg, 2.02 mmol) was added, followed bydiisopropylethylamine (0.34 mL, 2.02 mmol) and the reaction was stirredfor 3 hours at room temperature. Ethylamine hydrochloride-d₅ (24a) (165mg, 2.02 mmol, Sigma Aldrich, 99% D) was added, followed immediately bydiisopropylethylamine (0.34 mL, 2.02 mmol) and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered and concentratedunder reduced pressure. The residual material was purified using anAnalogix automated chromatography system eluting with methanol indichloromethane (0 to 5%). Product fractions were pooled and evaporatedto give a material with enantiomeric purity of 88% ee (350 mg). Thematerial was further purified using chiral preparative HPLC (20×250 mm,10 μm column, Daicel ChiralPak AD, eluting with 80% isopropanol/20%hexane at a flow rate of 17 mL/min). Product fractions were pooled andevaporated to afford (101) as an off white solid (210 mg, 48%). ¹H-NMR(300 MHz, CDCl₃): δ 2.61 (s, 3H), 3.29-3.36 (m, 1H), 3.47-3.54 (m, 1H),3.80 (s, 3), 4.61 (t, J=7 Hz, 1H), 6.35 (bm, 1H), 6.85-6.86 (d, J=2.9Hz, 1H), 7.17-7.21 (m, 1H), 7.22-7.34 (m, 2H), 7.36, (s, 1H), 7.43-7.5(m, 2H); ¹³C-NMR (75 MHz, CDCl₃): δ 12.13, 39.49, 53.93, 55.87, 115.81,117.94, 124.80, 126.46, 128.50, 130.11, 130.73, 136.95, 137.14, 150.46,156.42, 157.98, 166.19, 170.32; MS(ESI) [(M+H)⁺] C₂₂H₁₇D₅ClN₅O₂: 429;HPLC (method: SorbTech C18AQ, 2.1×50 mm 3 μm column-gradient method5-95% ACN+0.1% formic acid in 14 min with 4 min hold at 95% ACN+0.1%formic acid; Wavelength: 254 nm): retention time: 6.341 min; 98.6%purity; Chiral HPLC (method: 25 cm×4.6 mm, 10 μm column Chiralpak AD,isocratic method 40% heptane +60% EtOH for 30 minutes at 1.0 mL/min,Wavelength: 210 nm): retention time: 4.650 min, purity: 99.14% ee.

Example 4. (S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d₃)-1-(methyl-d₃)-4Hbenzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 112)

Step 1. Methyl(S,Z)-2-(2-(2-(acetyl-d₃)hydrazono)-5-(4-chlorophenyl)-7-(methoxy-d₃)-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)acetate(18c)

To a solution of 17a (1.2 g, 3.07 mmol) in THF (50 mL) at 0° C. wasadded Hydrazine monohydrate (0.44 mL, 9.3 mmol). The reaction wasstirred at 0° C. for 4 hours and triethylamine (1.3 mL, 9.3 mmol) wasadded, followed by acetyl chloride-d₃ (23a) (0.7 mL, 9.3 mmol, CambridgeIsotopes, 98% D). The reaction was stirred at room temperature for 1hour, diluted with water (50 mL) and THF was removed under reducedpressure. The resulting residue was extracted with dichloromethane (100mL), washed with saturated sodium chloride (100 mL), dried over sodiumsulfate, filtered, and concentrated to give crude titled compound (18c)as a brown solid (1.5 g, 99%).

Step 2. Methyl(S)-2-(6-(4-chlorophenyl)-8-(methoxy-d₃)-1-(methyl-d₃)-4H-benzo[f][1,2,4]-triazolo[4,3-a][1,4]diazepin-4-yl)acetate(19c)

To a solution of 18c (1.5 g, 3.07 mmol) in THF (40 mL) was added aceticacid (40 mL) and the reaction was stirred at room temperature for 18 h.The mixture was concentrated under reduced pressure to afford a residuewhich was dissolved in dichloromethane (150 mL), washed with saturatedsodium bicarbonate solution (3×50 mL), water (100 mL) and saturatedsodium chloride (100 mL), dried over sodium sulfate, filtered, andconcentrated under reduced pressure. The residual material was purifiedusing an Analogix automated chromatography system eluting with methanolin dichloromethane (0 to 5%). Product fractions were pooled andevaporated to afford 19c as a light yellow solid (1.1 g, 85%).

Step 3.(S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d₃)-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 112)

To a solution of 19c (1.1 g, 2.55 mmol) in THF (20 mL) was added 1N NaOH(7.7 mL, 7.7 mmol). The reaction was stirred for 5 hours at 40° C.,cooled to 0° C. and adjusted with 1N HCl solution to pH 4-5. Theresulting mixture was extracted with dichloromethane (3×50 mL) and thecombined organic layer was dried over sodium sulfate, filtered, andconcentrated to give the free acid as a yellow solid (960 mg, 93%). To asolution of the free acid (480 mg, 1.17 mmol) in THF (20 mL) was addedHATU (880 mg, 2.34 mmol), followed by diisopropylethylamine (0.4 mL,2.34 mmol) and the reaction was stirred at room temperature for 3 hours.Ethylamine hydrochloride (24) (190 mg, 2.34 mmol) was added, followedimmediately by diisopropylethylamine (0.4 mL, 2.34 mmol) and thereaction was stirred at room temperature for 18 hours. The resultingmixture was concentrated under reduced pressure and the residualmaterial was dissolved in a mixture of water (30 mL) and dichloromethane(30 mL), stirred for 30 minutes and extracted with dichloromethane (2×50mL). The combined organic phase was washed with water (2×100 mL),saturated sodium chloride (100 mL), dried over sodium sulfate, filteredand concentrated. The crude residue was purified using an Analogixautomated chromatography system eluting with methanol in dichloromethane(0 to 5%). Product fractions were pooled and evaporated to give amaterial with enantiomeric purity of 92% ee (390 mg). The material wasfurther purified using chiral preparative HPLC (method: Daicel ChiralPakAD 20×250 mm, 10 Lm column, eluting with 80% isopropanol/20% hexane at aflow rate of 17 mL/min). Product fractions were pooled and evaporated toafford 112 as an off white solid (240 mg, 49%). ¹H-NMR (300 MHz, CDCl₃):δ 1.18 (m, 3H), 3.23-3.32 (m, 2H), 3.34-3.40 (m, 1H), 3.41-3.54 (m, 1H),4.62 (t, J=7 Hz, 1H), 6.39 (bm, 1H), 6.85 (d, J=2.9 Hz, 1H), 7.17-7.21(m, 1H), 7.32-7.34 (m, 2H), 7.35, (s, 1H), 7.46-7.5 (m, 2H); ¹³C-NMR (75MHz, CDCl₃): δ 14.78, 34.49, 39.49, 53.93, 115.79, 117.94, 124.79,126.45, 128.50, 130.11, 130.74, 136.95, 137.15, 156.43, 157.97, 166.19,170.30. MS(ESI) [(M+H)⁺] C₂₂H₁₆D₆ClN₅O₂: 430; HPLC (method: SorbTechC18AQ, 2.1×50 mm 3 μm column-gradient method 5-95% ACN+0.1% formic acidin 14 min with 4 min hold at 95% ACN+0.1% formic acid; Wavelength: 254nm): retention time: 6.314 min; 99.2% purity; Chiral HPLC (method: 25cm×4.6 mm, 10 μm column Chiralpak AD, isocratic method 40% heptane +60%EtOH for 30 minutes at 1.0 mL/min, Wavelength: 210 nm): retention time:4.652 min, purity: 98.06% ee.

Example 5.(S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d₃)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d5)acetamide(Compound 113)

(S)-2-(6-(4-Chlorophenyl)-8-(methoxy-d₃)-1-methyl-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d5)acetamide(Compound 113)

To a solution of the free acid of 19a (500 mg, 1.25 mmol) (prepared bytreating with 1N NaOH/THF as described in step 3 of Compound 112) in THF(20 mL) was added HATU (950 mg, 2.5 mmol) followed bydiisopropylethylamine (0.42 mL, 2.5 mmol) and the reaction was stirredfor 3 hours at room temperature. Ethylamine hydrochloride-d₅ (24a) (220mg, 2.5 mmol, Sigma Aldrich, 99% D) was added followed bydiisopropylethylamine (0.42 mL, 2.5 mmol) and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL) and saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The resulting residue was purified using anAnalogix automated chromatography system eluting with methanol indichloromethane (0 to 5%). Product fractions were pooled and evaporatedto give a material with enantiomeric purity of 94% ee (400 mg). Thematerial was further purified using chiral preparative HPLC (DaicelChiralPak AD 20×250 mm, 10 μm column, eluting with 80% isopropanol/20%hexane at a flow rate of 17 mL/min). Product fractions were pooled andevaporated to afford 113 as an off white solid (280 mg, 51%). ¹H-NMR(300 MHz, CDCl₃): δ 2.61 (s, 3H), 3.29-3.36 (m, 1H), 3.47-3.54 (m, 1H),4.61 (t, J=7 Hz, 1H), 6.36 (bm, 1H), 6.85 (d, J=2.9 Hz, 1H), 7.17-7.21(m, 1H), 7.32-7.35 (m, 2H), 7.36, (s, 1H), 7.46-7.5 (m, 2H); ¹³C-NMR (75MHz, CDCl₃): δ 12.13, 39.49, 115.79, 117.92, 124.81, 126.44, 128.49,130.12, 130.73, 136.94, 137.15, 150.46, 156.43, 157.98, 166.19, 170.33.MS(ESI) [(M+H)⁺] C₂₂H₁₄D₈ClN₅O₂: 432; HPLC (method: SorbTech C18AQ,2.1×50 mm 3 μm column-gradient method 5-95% ACN+0.1% formic acid in 14min with 4 min hold at 95% ACN+0.1% formic acid; Wavelength: 254 nm):retention time: 6.301 min; 99.1% purity; Chiral HPLC (method: 25 cm×4.6mm, 10 μm column Chiralpak AD, isocratic method 40% heptane +60% EtOHfor 30 minutes at 1.0 mL/min, Wavelength: 210 nm): retention time: 4.645min, purity: 98.90% ee.

Example 6.(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d₅)acetamide(Compound 116)

(S)-2-(6-(4-chlorophenyl)-8-methoxy-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-(ethyl-d₅)acetamide(Compound 116)

To a solution of the free acid of 19b (500 mg, 1.4 mmol) (prepared bytreating with 1N NaOH/THF as described in step 1 of Compound 113) in THF(20 mL) was added HATU (1.06 g, 2.8 mmol) followed bydiisopropylethylamine (0.48 mL, 2.8 mmol) and the reaction was stirredat room temperature for 3 hours. Ethylamine hydrochloride-d₅ (24a) (240mg, 2.8 mmol, Sigma Aldrich, 99% D) was added, followed by anddiisopropylethylamine (0.48 mL, 2.5 mmol), and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered and concentrated.The resulting residue was purified using an Analogix automatedchromatography system eluting with methanol in dichloromethane (0 to5%). Product fractions were pooled and evaporated to give a materialwith enantiomeric purity of 88% ee (400 mg). The material was furtherpurified using chiral preparative HPLC (Daicel ChiralPak AD, 20×250 mm,10 μm column, eluting with 80% isopropanol/20% hexane at a flow rate of17 mL/min). Product fractions were pooled and evaporated to afford 116as an off white solid (320 mg, 57%). ¹H-NMR (300 MHz, CDCl₃): δ3.29-3.36 (m, 1H), 3.47-3.54 (m, 1H), 3.8 (s, 3H), 4.61 (t, J=7 Hz, 1H),6.35 (bm, 1H), 6.86 (d, J=2.9 Hz, 1H), 7.18-7.21 (m, 1H), 7.31-7.35 (m,2H), 7.36, (s, 1H), 7.46-7.51 (m, 2H); ¹³C-NMR (75 MHz, CDCl₃): δ 39.47,53.92, 55.87, 115.81, 117.94, 124.78, 126.48, 128.49, 130.11, 130.73,136.94, 137.15, 156.43, 157.97, 162.61, 166.18, 170.33. MS(ESI) [(M+H)⁺]C₂₂H₁₄D₈ClN₅O₂: 432; HPLC (method: SorbTech C18AQ, 2.1×50 mm 3 μmcolumn-gradient method 5-95% ACN+0.1% formic acid in 14 min with 4 minhold at 95% ACN+0.1% formic acid; Wavelength: 254 nm): retention time:6.343 min; 98.6% purity; Chiral HPLC (method: 25 cm×4.6 mm, 10 μm columnChiralpak AD, isocratic method 40% heptane +60% EtOH for 30 minutes at1.0 mL/min, Wavelength: 210 nm): retention time: 4.643 min, purity:97.87% ee.

Example 7.(S)-2-(6-(4-chlorophenyl)-8-(methoxy-d₃)-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 126)

(S)-2-(6-(4-chlorophenyl)-8-(methoxy-d₃)-1-(methyl-d₃)-4H-benzo[f][1,2,4]triazolo[4,3-a][1,4]diazepin-4-yl)-N-ethylacetamide(Compound 126)

To a solution of free acid of 19c (480 mg, 1.17 mmol) (prepared bytreating with 1N NaOH/THF as described in step 1 of 116) in THF (20 mL)was added HATU (880 mg, 2.34 mmol, 2.0 equiv) followed bydiisopropylethylamine (0.4 mL, 2.34 mmol) and the reaction was stirredat room temperature for 3 hours. Ethylamine hydrochloride-d₅ (24a) (200mg, 2.34 mmo, Sigma Aldrich, 99% D) was added, followed bydiisopropylethylamine (0.4 mL, 2.34 mmol) and the reaction was stirredat room temperature for 18 hours. The resulting mixture was concentratedunder reduced pressure and the residual material was dissolved in amixture of water (30 mL) and dichloromethane (30 mL), stirred for 30minutes and extracted with dichloromethane (2×50 mL). The combinedorganic phase was washed with water (2×100 mL), saturated sodiumchloride (100 mL), dried over sodium sulfate, filtered and concentrated.The resulting residue was purified using an Analogix automatedchromatography system eluting with methanol in dichloromethane (0 to5%). Product fractions were pooled and evaporated to give a materialwith enantiomeric purity of 92% ee (380 mg). The material was furtherpurified using chiral preparative HPLC (Daicel ChiralPak AD 20×250 mm,10 μm column, eluting with 80% isopropanol/20% hexane at a flow rate of17 mL/min). Product fractions were pooled and evaporated to afford 126as an off white solid (240 mg, 49%). ¹H-NMR (300 MHz, CDCl₃): δ3.29-3.36 (m, 1H), 3.47-3.54 (m, 1H), 4.62 (t, J=7 Hz, 1H), 6.41 (bs,1H), 6.85 (d, J=2.9 Hz, 1H), 7.18-7.22 (m, 1H), 7.27-7.32 (m, 2H), 7.36,(s, 1H), 7.47-7.51 (m, 2H); ¹³C-NMR (75 MHz, CDCl₃): δ 39.46, 53.92,115.78, 117.96, 124.81, 126.41, 128.52, 130.08, 130.75, 136.94, 137.14,156.41, 157.97, 166.27, 170.35. MS(ESI) [(M+H)⁺] C₂₂H₁₁D₁₁ClN₅O₂: 435;HPLC (method: SorbTech C18AQ, 2.1×50 mm 3 μm column-gradient method5-95% ACN+0.1% formic acid in 14 min with 4 min hold at 95% ACN+0.1%formic acid; Wavelength: 254 nm): retention time: 6.325 min; 98.6%purity; Chiral HPLC (method: 25 cm×4.6 mm, 10 μm column Chiralpak AD,isocratic method 40% heptane +60% EtOH for 30 minutes at 1.0 mL/min,Wavelength: 210 nm): retention time: 4.644 min, purity: 100% ee.

Example X. Evaluation of Metabolic Stability

Microsomal Assay:

Human liver microsomes (20 mg/mL) are obtained from Xenotech, LLC(Lenexa, Kans.). β-nicotinamide adenine dinucleotide phosphate, reducedform (NADPH), magnesium chloride (MgCl₂), and dimethyl sulfoxide (DMSO)are purchased from Sigma-Aldrich.

Determination of Metabolic Stability:

7.5 mM stock solutions of test compounds are prepared in DMSO. The 7.5mM stock solutions are diluted to 12.5-50 μM in acetonitrile (ACN). The20 mg/mL human liver microsomes are diluted to 0.625 mg/mL in 0.1 Mpotassium phosphate buffer, pH 7.4, containing 3 mM MgCl₂. The dilutedmicrosomes are added to wells of a 96-well deep-well polypropylene platein triplicate. A 10 μL aliquot of the 12.5-50 μM test compound is addedto the microsomes and the mixture is pre-warmed for 10 minutes.Reactions are initiated by addition of pre-warmed NADPH solution. Thefinal reaction volume is 0.5 mL and contains 0.5 mg/mL human livermicrosomes, 0.25-1.0 μM test compound, and 2 mM NADPH in 0.1 M potassiumphosphate buffer, pH 7.4, and 3 mM MgCl₂. The reaction mixtures areincubated at 37° C., and 50 μL aliquots are removed at 0, 5, 10, 20, and30 minutes and added to shallow-well 96-well plates which contain 50 μLof ice-cold ACN with internal standard to stop the reactions. The platesare stored at 4° C. for 20 minutes after which 100 μL of water is addedto the wells of the plate before centrifugation to pellet precipitatedproteins. Supernatants are transferred to another 96-well plate andanalyzed for amounts of parent remaining by LC-MS/MS using an AppliedBio-systems API 4000 mass spectrometer. The same procedure is followedfor the non-deuterated counterpart of the compound of Formula I and thepositive control, 7-ethoxycoumarin (1 μM). Testing is done intriplicate.

Data analysis: The in vitro t_(1/2)s for test compounds are calculatedfrom the slopes of the linear regression of % parent remaining (ln) vsincubation time relationship.in vitro t _(1/2)=0.693/kk=−[slope of linear regression of % parent remaining(ln) vs incubationtime]

Data analysis is performed using Microsoft Excel Software.

Without further description, it is believed that one of ordinary skillin the art can, using the preceding description and the illustrativeexamples, make and utilize the compounds of the present invention andpractice the claimed methods. It should be understood that the foregoingdiscussion and examples merely present a detailed description of certainpreferred embodiments. It will be apparent to those of ordinary skill inthe art that various modifications and equivalents can be made withoutdeparting from the spirit and scope of the invention.

I claim:
 1. A method of treating a subject suffering from a chronicautoimmune or inflammatory condition selected from rheumatoid arthritis,osteoarthritis, acute gout, psoriasis, systemic lupus erythematosus,multiple sclerosis, inflammatory bowel disease, Crohn's diseaseulcerative colitis, asthma, chronic obstructive airways disease,pneumonitis, myocarditis, pericarditis, myositis, eczema, dermatitis,alopecia, vitiligo, bullous skin diseases, nephritis, vasculitis,atherosclerosis, Alzheimer's disease, depression, retinitis, uveitis,scleritis, hepatitis, pancreatitis, primary biliary cirrhosis,sclerosing cholangitis, Addison's disease, hypophysitis, thyroiditis,type I diabetes and acute rejection of transplanted organs, the methodcomprising a step of administering to the subject in need thereof acompound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein Y^(1a) and Y^(1b)are the same and are hydrogen or deuterium; Y² is hydrogen or deuterium;R¹ and R² are each independently selected from CH₃ and CD₃; R³ isselected from CH₂CH₃, CH₂CD₃, CD₂CH₃, and CD₂CD₃; and if R¹ and R² areeach CH₃, R³ is CH₂CH₃, and Y² is hydrogen, then Y^(1a) and Y^(1b) areeach deuterium; and wherein any atom not designated as deuterium ispresent at its natural isotopic abundance.
 2. The method of claim 1,wherein the inflammatory condition is dermatitis.
 3. The method of claim1, wherein the inflammatory bowel disease is Crohn's disease orulcerative colitis.
 4. The method of claim 1, wherein for the compoundof Formula I, Y^(1a) and Y^(1b) are hydrogen.
 5. The method of claim 1,wherein for the compound of Formula I, Y^(1a) and Y^(1b) are deuterium.6. The method of claim 1, wherein for the compound of Formula I, R³ isselected from CH₂CH₃ and CD₂CD₃.
 7. The method of claim 4, wherein thecompound of Formula I is selected from any one of the compounds setforth in the table below: Compound Y² R¹ R² R³ 101 H —CH₃ —CH₃ —CD₂CD₃102 H —CH₃ —CH₃ —CD₂CH₃ 103 H —CH₃ —CH₃ —CH₂CD₃ 104 H —CD₃ —CH₃ —CH₂CH₃105 H —CH₃ —CD₃ —CH₂CH₃ 106 D —CH₃ —CH₃ —CH₂CH₃ 107 D —CD₃ —CH₃ —CH₂CH₃108 D —CH₃ —CD₃ —CH₂CH₃ 109 D —CH₃ —CH₃ —CD₂CD₃ 110 D —CH₃ —CH₃ —CD₂CH₃111 D —CH₃ —CH₃ —CH₂CD₃ 112 H —CD₃ —CD₃ —CH₂CH₃ 113 H —CD₃ —CH₃ —CD₂CD₃114 H —CD₃ —CH₃ —CD₂CH₃ 115 H —CD₃ —CH₃ —CH₂CD₃ 116 H —CH₃ —CD₃ —CD₂CD₃117 H —CH₃ —CD₃ —CD₂CH₃ 118 H —CH₃ —CD₃ —CH₂CD₃ 119 D —CD₃ —CD₃ —CH₂CH₃120 D —CD₃ —CH₃ —CD₂CD₃ 121 D —CD₃ —CH₃ —CD₂CH₃ 122 D —CD₃ —CH₃ —CH₂CD₃123 D —CH₃ —CD₃ —CD₂CD₃ 124 D —CH₃ —CD₃ —CD₂CH₃ 125 D —CH₃ —CD₃ —CH₂CD₃126 H —CD₃ —CD₃ —CD₂CD₃ 127 H —CD₃ —CD₃ —CD₂CH₃ 128 H —CD₃ —CD₃ —CH₂CD₃129 D —CD₃ —CD₃ —CD₂CD₃ 130 D —CD₃ —CD₃ —CD₂CH₃ 131 D —CD₃ —CD₃ —CH₂CD₃

or a pharmaceutically acceptable salt thereof.
 8. The method of claim 5,wherein the compound of Formula I is selected from any one of thecompounds set forth in the table below: Compound Y² R¹ R² R³ 200 H —CH₃—CH₃ —CH₂CH₃ 201 H —CH₃ —CH₃ —CD₂CD₃ 202 H —CH₃ —CH₃ —CD₂CH₃ 203 H —CH₃—CH₃ —CH₂CD₃ 204 H —CD₃ —CH₃ —CH₂CH₃ 205 H —CH₃ —CD₃ —CH₂CH₃ 206 D —CH₃—CH₃ —CH₂CH₃ 207 D —CD₃ —CH₃ —CH₂CH₃ 208 D —CH₃ —CD₃ —CH₂CH₃ 209 D —CH₃—CH₃ —CD₂CD₃ 210 D —CH₃ —CH₃ —CD₂CH₃ 211 D —CH₃ —CH₃ —CH₂CD₃ 212 H —CD₃—CD₃ —CH₂CH₃ 213 H —CD₃ —CH₃ —CD₂CD₃ 214 H —CD₃ —CH₃ —CD₂CH₃ 215 H —CD₃—CH₃ —CH₂CD₃ 216 H —CH₃ —CD₃ —CD₂CD₃ 217 H —CH₃ —CD₃ —CD₂CH₃ 218 H —CH₃—CD₃ —CH₂CD₃ 219 D —CD₃ —CD₃ —CH₂CH₃ 220 D —CD₃ —CH₃ —CD₂CD₃ 221 D —CD₃—CH₃ —CD₂CH₃ 222 D —CD₃ —CH₃ —CH₂CD₃ 223 D —CH₃ —CD₃ —CD₂CD₃ 224 D —CH₃—CD₃ —CD₂CH₃ 225 D —CH₃ —CD₃ —CH₂CD₃ 226 H —CD₃ —CD₃ —CD₂CD₃ 227 H —CD₃—CD₃ —CD₂CH₃ 228 H —CD₃ —CD₃ —CH₂CD₃ 229 D —CD₃ —CD₃ —CD₂CD₃ 230 D —CD₃—CD₃ —CD₂CH₃ 231 D —CD₃ —CD₃ —CH₂CD₃

or a pharmaceutically acceptable salt thereof.
 9. The method of claim 1,wherein for the compound of Formula I, the compound has an isotopicenrichment factor for each designated deuterium atom of at least 6000(90% deuterium incorporation).
 10. The method of claim 1, wherein forthe compound of Formula I, the compound has an isotopic enrichmentfactor for each designated deuterium atom of at least 6333.3 (95%deuterium incorporation).
 11. The method of claim 7, wherein thecompound of Formula I is selected from any one of the compounds setforth in the table below: Compound Y² R¹ R² R³ 101 H —CH₃ —CH₃ —CD₂CD₃105 H —CH₃ —CD₃ —CH₂CH₃ 108 D —CH₃ —CD₃ —CH₂CH₃ 109 D —CH₃ —CH₃ —CD₂CD₃112 H —CD₃ —CD₃ —CH₂CH₃ 113 H —CD₃ —CH₃ —CD₂CD₃ 116 H —CH₃ —CD₃ —CD₂CD₃117 H —CH₃ —CD₃ —CD₂CH₃ 118 H —CH₃ —CD₃ —CH₂CD₃ 119 D —CD₃ —CD₃ —CH₂CH₃120 D —CD₃ —CH₃ —CD₂CD₃ 123 D —CH₃ —CD₃ —CD₂CD₃ 124 D —CH₃ —CD₃ —CD₂CH₃125 D —CH₃ —CD₃ —CH₂CD₃ 126 H —CD₃ —CD₃ —CD₂CD₃ 127 H —CD₃ —CD₃ —CD₂CH₃128 H —CD₃ —CD₃ —CH₂CD₃ 129 D —CD₃ —CD₃ —CD₂CD₃ 130 D —CD₃ —CD₃ —CD₂CH₃131 D —CD₃ —CD₃ —CH₂CD₃

or a pharmaceutically acceptable salt thereof, wherein the compound hasan isotopic enrichment factor for each designated deuterium atom of atleast 6000 (90% deuterium incorporation).
 12. The method of claim 8,wherein the compound of Formula I is selected from any one of thecompounds set forth in the table below: Compound Y² R¹ R² R³ 201 H —CH₃—CH₃ —CD₂CD₃ 205 H —CH₃ —CD₃ —CH₂CH₃ 208 D —CH₃ —CD₃ —CH₂CH₃ 209 D —CH₃—CH₃ —CD₂CD₃ 212 H —CD₃ —CD₃ —CH₂CH₃ 213 H —CD₃ —CH₃ —CD₂CD₃ 216 H —CH₃—CD₃ —CD₂CD₃ 217 H —CH₃ —CD₃ —CD₂CH₃ 218 H —CH₃ —CD₃ —CH₂CD₃ 219 D —CD₃—CD₃ —CH₂CH₃ 220 D —CD₃ —CH₃ —CD₂CD₃ 223 D —CH₃ —CD₃ —CD₂CD₃ 224 D —CH₃—CD₃ —CD₂CH₃ 225 D —CH₃ —CD₃ —CH₂CD₃ 226 H —CD₃ —CD₃ —CD₂CD₃ 227 H —CD₃—CD₃ —CD₂CH₃ 228 H —CD₃ —CD₃ —CH₂CD₃ 229 D —CD₃ —CD₃ —CD₂CD₃ 230 D —CD₃—CD₃ —CD₂CH₃ 231 D —CD₃ —CD₃ —CH₂CD₃

or a pharmaceutically acceptable salt thereof, wherein the compound hasan isotopic enrichment factor for each designated deuterium atom of atleast 6000 (90% deuterium incorporation).
 13. The method of claim 11,wherein for the compound of Formula I, the compound has an isotopicenrichment factor for each designated deuterium atom of at least 6333.3(95% deuterium incorporation).
 14. The method of claim 12, wherein forthe compound of Formula I, the compound has an isotopic enrichmentfactor for each designated deuterium atom of at least 6333.3 (95%deuterium incorporation).